减体积肝移植模型大鼠肝脏组织miRNAs表达谱变化  

Changes in miRNAs expression profiling in the liver of a rat model after reduced-size liver transplantation

作　　者：高红强[1] 李志强[1] 王海富[1] 薛国友[1] 刘静[1] 陈刚[1] 李立[1] 

机构地区：[1]昆明医科大学附属甘美医院昆明市第一人民医院肝胆胰外科,云南省昆明市650011

出　　处：《中国组织工程研究》2016年第5期712-717,共6页Chinese Journal of Tissue Engineering Research

基　　金：云南省应用基础研究计划-昆明医科大学联合专项重点项目(2012FB010)~~

摘　　要：背景:活体肝移植后肝脏再生是影响肝移植预后的关键因素,目前尚未见调控活体肝移植术后肝脏再生的特异性微小RNA(miRNA)的报道。目的:分析miRNAs在大鼠减体积肝移植后各时间点表达谱的变化,挑选并验证目的 miRNAs,为大鼠减体积肝移植后肝脏再生干预策略提供靶标miRNAs,为临床活体肝移植后肝脏再生研究提供理论依据。方法:分别建立大鼠减体积肝移植模型和假手术模型,利用miRNAs基因芯片技术检测miRNAs表达谱的差异。在差异表达的miRNAs中挑选出目的 miRNAs进行实时定量PCR检测,验证芯片结果的可信性。结果与结论:与假手术组大鼠肝组织相比,大鼠减体积肝移植肝脏组织中表达上调的miRNAs有11个,分别为let-7b-5p,let-7c-5p,miR-101a-3p,miR-103-3p,miR-130a-3p,miR-142-5p,miR-186-5p,miR-199a-3p,miR-21-5p,221-3p,miR-34a-5p;表达下调的miRNAs有4个miR-26b-5p,miR-150-5p,miR-19a-3p,rno-miR-146-5p。将挑选出的目的 miRNAs进行PCR验证发现,miR-221-3p,miR-199a-3p在24 h,48 h,1周表达量与所对应的芯片结果近似,各自变化趋势与芯片结果一致,说明miRNA芯片结果可信。结果证实,大鼠减体积肝移植后伴有miRNAs表达谱的变化,实验挑选并验证了目的 miRNAs。BACKGROUND： Liver regeneration is the key factor influencing the prognosis of living donor liver transplantation. There has not been the research on special miRNA of liver regeneration after living donor liver transplantation. OBJECTIVE： To analyze the variation of miRNAs expression profile after rat reduced-size liver transplantation at certain time point, select and verify target miRNA which can provide targeting intervention strategies in liver regeneration after rat reduced-size liver transplantation and provide theoretical evidence for liver regeneration after living donor liver transplantation.METHODS： The reduced-size liver transplantation models were established. miRNAs microarray was used to detect miRNA expression. In differentially expressed micro RNAs, real-time quantitative PCR was utilized to detect target miRNAs. The credibility of miRNAs microarray results was verified. RESULTS AND CONCLUSION： Compared with rat liver tissue in the sham operation group, 11 miRNAs up-regulated in reduced-size liver transplantation, including let-7b-5p, let-7c-5p, miR-101a-3p, miR-103-3p, miR-130a-3p, miR-142-5p, miR-186-5p, miR-199a-3p, miR-21-5p, 221-3p and miR-34a-5p. Four miRNAs were down-regulated, including miR-26b-5p, miR-150-5p, miR-19a-3p and rno-miR-146-5p. PCR test further verified that miR-221-3p and miR-199a-3p expression changes approximated the chip results at 24, 48 hours and 1 week, indicating that results of miRNA microarray were believable. These results verified that it exists variation of miRNAs expression profile after rat reduced-size liver transplantation, which picked out and verified the target miRNAs.

关 键 词：组织工程 肝移植 芯片 实验动物 移植动物模型 大鼠减体积肝移植 消化系统损伤模型 肝脏 微小RNA MIRNAS MICROARRAY PCR 反转录 miRNAs表达谱 miRNAs芯片数据分析 

分 类 号：R318[医药卫生—生物医学工程]